增强型靶向NGS检测在结核病和耐药结核病患者综合诊断中的应用

IF 4.6 2区医学 Q1 INFECTIOUS DISEASES

International Journal of Antimicrobial Agents Pub Date : 2025-08-20 DOI:10.1016/j.ijantimicag.2025.107598

Miaoran Wang , Ying Wu , Ying Xi , Lichao Fan , Fengshuo Sun , Gang Zhou , Wei Wang , Yanhong Yu , Jiao Sun , Xue Yang , Jun Tang , Lifeng Li , Yu Chen

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引用次数: 0

摘要

背景：准确和及时诊断结核病和耐药结核病至关重要；然而，目前的方法在灵敏度和范围上存在局限性，特别是在检测耐药性和区分其他呼吸道病原体方面。方法：建立了一种基于多重pcr的靶向下一代测序（tNGS）分析方法，并验证了其分析性能，包括检出限（LoD）、精密度和抗干扰性。对181名疑似结核病患者进行了一项单中心前瞻性研究，通过支气管肺泡灌洗或痰样本评估该检测方法的临床性能，并与微生物培养和Xpert MTB/RIF进行比较。结果：tNGS法检测结核及其他呼吸道病原菌的LoD低，为10拷贝/mL，检测精度高，抗干扰能力强。在临床验证中，它在结核病确诊病例中达到了94.94%的敏感性，在利福平耐药性方面与Xpert MTB/RIF的阳性率为92.86%，同时还识别了其他突变。该方法可准确检测出非结核分枝杆菌（NTM）和其他呼吸道病原体，有助于鉴别诊断。结论：tNGS检测可可靠地检测结核病、耐药突变和包括NTM在内的呼吸道病原体，从而加强疑似结核病患者的鉴别诊断并支持有效的治疗策略。

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Enhanced Targeted NGS Assay for Comprehensive Diagnosis in Tuberculosis and Drug-Resistant Tuberculosis Patients

Background

Accurate and timely diagnosis of tuberculosis (TB) and drug-resistant TB is crucial; however, current methods have limitations in sensitivity and scope, especially for detecting drug resistance and differentiating other respiratory pathogens.

Methods

We developed a multiplex PCR-based targeted next-generation sequencing (tNGS) assay and validated its analytical performance, including limit of detection (LoD), precision, and resistance to interference. A single-centre prospective study was conducted with 181 suspected TB patients to evaluate the assay’s clinical performance using bronchoalveolar lavage or sputum samples, compared against microbiological culture and Xpert MTB/RIF.

Results

The tNGS assay demonstrated a low LoD of 10 copies/mL for TB and other respiratory pathogens, with high precision and resistance to interference. In clinical validation, it achieved 94.94 % sensitivity in confirmed TB cases and showed a 92.86 % positive percent agreement with Xpert MTB/RIF for rifampicin resistance, while also identifying additional mutations. The assay accurately detected nontuberculous mycobacteria (NTM) and other respiratory pathogens, aiding differential diagnosis.

Conclusion

The tNGS assay provides reliable detection of TB, drug-resistant mutations, and respiratory pathogens, including NTM, thereby enhancing differential diagnosis and supporting effective treatment strategies in the management of patients with suspected TB.

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来源期刊

International Journal of Antimicrobial Agents 医学-传染病学

CiteScore

21.60

自引率

0.90%

发文量

176

审稿时长

36 days

期刊介绍： The International Journal of Antimicrobial Agents is a peer-reviewed publication offering comprehensive and current reference information on the physical, pharmacological, in vitro, and clinical properties of individual antimicrobial agents, covering antiviral, antiparasitic, antibacterial, and antifungal agents. The journal not only communicates new trends and developments through authoritative review articles but also addresses the critical issue of antimicrobial resistance, both in hospital and community settings. Published content includes solicited reviews by leading experts and high-quality original research papers in the specified fields.